Benzothiophenes, formulations containing same, and methods

ABSTRACT

This invention provides compounds of formula I 
     
       
         
         
             
             
         
       
     
     and pharmaceutically acceptable salts and solvates thereof, characterized that the compound is in particulate form and having a specific size range. 
     The present invention further provides pharmaceutical compositions containing or formulated using compounds of formula I, and the use of such compounds for alleviating human pathologies, including osteoporosis, serum lipid lowering, and breast cancer.

FIELD OF THE INVENTION

This invention relates to the fields of pharmaceutical and organicchemistry and provides a benzothiophene compound, in particulate form,which is useful for the treatment of various medical indications,including osteoporosis and lipid lowering. More particularly, thebenzothiophene is of a particle size range which allows enhancedbioavailabilty and control during the manufacturing process.

BACKGROUND OF THE INVENTION

Osteoporosis describes a group of diseases which arise from diverseetiologies, but which are characterized by the net loss of bone mass perunit volume. The consequence of this loss of bone mass and resultingbone fracture is the failure of the skeleton to provide adequatestructural support for the body. One of the most common types ofosteoporosis is that associated with menopause. Most women lose fromabout 20% to about 60% of the bone mass in the trabecular compartment ofthe bone within 3 to 6 years after the cessation of menses. This rapidloss is generally associated with an increase of bone resorption andformation. However, the resorptive cycle is more dominant and the resultis a net loss of bone mass. Osteoporosis is a common and serious diseaseamong post-menopausal women.

There are an estimated 25 million women in the United States, alone, whoare afflicted with this disease. The results of osteoporosis arepersonally harmful and also account for a large economic loss due itschronicity and the need for extensive and long term support(hospitalization and nursing home care) from the disease sequelae. Thisis especially true in more elderly patients. Additionally, althoughosteoporosis is not generally thought of as a life threateningcondition, a 20% to 30% mortality rate is related with hip fractures inelderly women. A large percentage of this mortality rate can be directlyassociated with post-menopausal osteoporosis.

At this time, the only generally accepted methods for treatment ofpost-menopausal osteoporosis are estrogen replacement therapy and theuse of the bisphosphonate alendronate. Although therapy is generallysuccessful, patient compliance with the therapy is relatively lowprimarily, due to undesirable side effects.

Throughout premenopausal time, most women have less incidence ofcardiovascular disease than age-matched men. Following menopause,however, the rate of cardiovascular disease in women slowly increases tomatch the rate seen in men. This loss of protection has been linked tothe loss of estrogen and, in particular, to the loss of estrogen'sability to regulate the levels of serum lipids. The nature of estrogen'sability to regulate serum lipids is not well understood, but evidence todate indicates that estrogen can up regulate the low density lipid (LDL)receptors in the liver to remove excess cholesterol. Additionally,estrogen appears to have some effect on the biosynthesis of cholesterol,and other beneficial effects on cardiovascular health.

It has been reported in the literature that post-menopausal women havingestrogen replacement therapy have a return of serum lipid levels toconcentrations to those of the pre-menopausal state. Thus, estrogenwould appear to be a reasonable treatment for this condition. However,the side-effects of estrogen replacement therapy are not acceptable tomany women, thus limiting the use of this therapy. An ideal therapy forthis condition would be an agent which would regulate the serum lipidlevel as does estrogen, but would be devoid of the side-effects andrisks associated with estrogen therapy.

Preclinical findings with a structurally distinct “anti-estrogen”,raloxifene, have demonstrated potential for improved selectivity ofestrogenic effects in target tissues. Similar to tamoxifen, raloxifenewas developed originally for treatment of breast cancer; however, thebenzothiophene nucleus of raloxifene represented a significantstructural deviation from the triphenylethylene nucleus of tamoxifen.Raloxifene binds with high affinity to the estrogen receptor, andinhibits estrogen-dependent proliferation in MCF-7 cells (human mammarytumor derived cell line) in cell culture. In vivo estrogen antagonistactivity of raloxifene was furthermore demonstrated incarcinogen-induced models of mammary tumors in rodents. Significantly,in uterine tissue raloxifene was more effective than tamoxifen as anantagonist of the uterotrophic response to estrogen in immature ratsand, in contrast to tamoxifen, raloxifene displayed only minimaluterotrophic response that was not dose-dependent in ovariectomized(OVX) rats. Thus, raloxifene is unique as an antagonist of the uterineestrogen receptor, in that it produces a nearly complete blockage ofuterotrophic responses of estrogen due to minimal agonist effect ofraloxifene in this tissue. Indeed, the ability of raloxifene toantagonize the uterine stimulatory effect of tamoxifen was recentlydemonstrated in OVX rats. Raloxifene is more properly characterized as aSelective Estrogen Receptor Modulator (SERM), due to its unique profile.

Raloxifene is now in Phase III clinical trials for osteoporosis.Indications thus far from these trials and other data, point toraloxifene's potential not only as an osteoporosis therapy, but also ofpotential use in lowering LDL (serum lipid) levels, inhibitingendometriosis and uterine fibrosis, and preventing breast cancer. Theadvancement of raloxifene has been somewhat hampered by its physicalcharacteristics, both as to bioavailability and in manufacturing. Forexample, it is generally insoluble, and this can adversely affect thebioavailability. Clearly, any improvement in the physicalcharacteristics of raloxifene, would potentially offer a more beneficialtherapy and enhanced manufacturing capability.

SUMMARY OF THE INVENTION

This invention provides a compound of formula I

and pharmaceutically acceptable salts and solvates thereof,characterized in that the compound is in particulate form, saidparticles having a mean particle size of less than about 25 microns, andpreferably between about 5 and about 20 microns.

Further, the present invention encompasses compounds of formula Iwherein at least 90% of the particles have a particle size of less thanabout 50 microns, and preferably less than about 35 microns.

The present invention further relates to pharmaceutical compositionscontaining or formulated using one or more compounds of formula I,optionally containing estrogen or progestin, and the use of suchcompounds, alone, or in combination with estrogen or progestin, foralleviating the symptoms of osteoporosis lowering lipid levels, andinhibiting endometriosis, uterine fibrosis, and breast cancer.

DETAILED DESCRIPTION OF THE INVENTION

It has now been found that by processing compounds of formula I, tobring their particle size within a specified narrow range,pharmaceutical compositions may be prepared which exhibit for theiractive ingredient both a consistent in vitro dissolution profile and invivo bioavailability. In addition to bringing about these desireddissolution/bioavailability characteristics, the control of particlesize to a narrow range has also resulted in significant improvements inmanufacturing capabilities.

The mean particle size of the compounds of formula I, as set out by theinvention, is less than about 25 microns, preferably between about 5 andabout 20 microns. Further, the invention encompasses formula I compoundswith at least 90% of the particles having a particle size of less thanabout 50 microns, preferably less than about 35 microns. Morepreferably, the mean particle size range is between about 5 and about 20microns, with at least 90% of the particles having a size of less thanabout 35 microns.

It will of course be understood by those familiar with comminutionprocess techniques that the limit set on the size of 90% or more of theparticles is a limitation to further distinguish the particulatecompounds of the invention from those exhibiting a broader sizedistribution, because of the wide variation in size encountered in allmatter reduced in size by a process of comminution or particle sizereduction, for example, by milling utilizing a variety of kinds ofmilling equipment now available, for example, hammer, pin or fluidenergy mills.

The invention also provides pharmaceutical compositions comprising orformulated using the said particulate compound of the invention and oneor more pharmaceutically-acceptable excipients or carriers.

The term “inhibit” is defined to include its generally accepted meaningwhich includes prohibiting, preventing, restraining, and lowering,stopping, or reversing progression or severity, and such action on aresultant symptom. As such the present invention includes both medicaltherapeutic and prophylactic administration, as appropriate.

The term “molar equivalents,” as used herein, refers to the number ofmoles of the boron trihalide reagent in relation to the number of molesof the starting benzothiophene compound. For example, three millimolesof boron trichloride reacted with one millimole of the benzothiophenecompound would represent three molar equivalents of boron trichloride.

The term “solvate” represents an aggregate that comprises one or moremolecules of the solute, such as a formula I compound, with a moleculeof solvent. Representative solvates are formed with methylene chloride,1,2-dichloroethane, chloroform, and 1,2,3-trichloropropane.

A desirable form of raloxifene hydrochloride is the non-solvatedcrystalline form described in UK published patent application GB2293382(A), published Mar. 27, 1996, filed Aug. 18, 1995, ApplicationNumber 9519028.6, incorporated herein by reference.

As used herein, the term “estrogen” includes steroidal compounds havingestrogenic activity such as, for example, 17β-estradiol, estrone,conjugated estrogen (Premarin®), equine estrogen, 17β-ethynyl estradiol,and the like. As used herein, the term “progestin” includes compoundshaving progestational activity such as, for example, progesterone,norethylnodrel, nongestrel, megestrol acetate, norethindrone, and thelike.

The term “mean particle size” is defined as equivalent sphericaldiameter as determined by laser light diffraction scattering.

Raloxifene's chemical name is6-hydroxy-2-(4-hydroxyphenyl)-3-[4-(2-piperidinoethoxy)benzoyl]benzo[b]-thiophene.“Raloxifene” also encompasses the salts and solvates thereof, with thehydrochloride salt being preferred.

Raloxifene is a nuclear regulatory molecule or second generationselective estrogen receptor modulator (SERM). Raloxifene has been shownto bind to the estrogen receptor and was originally thought to be amolecule whose function and pharmacology was that of a pureanti-estrogen in that it blocked the ability of estrogen to activateuterine tissue and estrogen dependent breast cancers. Indeed, raloxifenedoes block the action of estrogen in some cells; however in other celltypes, raloxifene activates the same genes as estrogen does and displaysthe same pharmacology, e.g., osteoporosis, hyperlipidemia. The uniqueprofile which raloxifene displays and differs from that of estrogen isnow thought to be due to the unique activation and/or suppression ofvarious gene functions by the raloxifene-estrogen receptor complex asopposed to the activation and/or suppression of genes by theestrogen-estrogen receptor complex. Therefore, although raloxifene andestrogen utilize and compete for the same receptor, the pharmacologicaloutcome from gene regulation of the two is not easily predicted and isunique to each.

The compounds of the current invention can be made according toestablished procedures, such as those detailed in U.S. Pat. Nos.4,133,814, 4,418,068, and 4,380,635, and European Patent Application95306050.6, Publication No. 0699672, Kjell, et al., filed Aug. 30, 1995,published Mar. 6, 1996, all of which are incorporated by referenceherein. In general, the process starts with a benzo[b]thiophene having a6-hydroxyl group and a 2-(4-hydroxyphenyl) group. The starting compoundis protected, acylated, and deprotected to form the formula I compounds.Examples of the preparation of such compounds are provided in the U.S.patents discussed above.

Also, the information disclosed in the published European PatentApplication number 0670162 A1, published on Sep. 6, 1995, isincorporated by reference.

A preferred synthesis is set out as follows. R⁴ is hydrogen or C₁-C₄alkoxy, R⁵ is hydrogen or C₁-C₄ alkyl, and R⁶ is chloro, bromo, orhydroxyl. HX is HCl or HBr.

The Formula II and III compounds, the starting materials for thecompounds of formula I, are prepared using standard synthetic organicmethods. The Formula II starting compound is readily obtained by asynthesis which is exemplified below in Preparation I and outlined inScheme I.

The Formula II compounds, wherein R⁴ and R⁵ are as defined above, areprepared by first reacting a 3-alkoxybenzenethiol with phenacyl or4′-alkoxyphenacyl bromide in the presence of a strong base. Suitablebases for this transformation include, but are not limited to, potassiumhydroxide and sodium hydroxide. The reaction is typically carried out inethanol or a mixture of water and ethanol at a temperature of about 0°C. to about 50° C. The next step is cyclization of thearylphenacylsulfide. The cyclization is conveniently carried out byheating the arylphenacylsulfide in polyphosphoric acid. The cyclizationis typically carried out at a temperature of about 80° C. to about 120°C., preferably between 85° C. and 90° C. The Formula II benzothiopheneis typically purified by recrystallization. For example, when R⁴ ismethoxy and R⁵ is methyl, the formula II compound may be recrystallizedfrom ethyl acetate.

The acylating agent for the present process, a Formula III compound, isprepared as shown in Scheme II, wherein the variables R⁶ and HX are asdefined above and R is C₁-C₄ alkyl.

Generally, a C₁-C₄ alkyl 4-hydroxybenzoate is alkylated with achloroethylamine in the presence of an inorganic base and the estergroup hydrolyzed to produce the Formula III compounds, wherein R⁶ ishydroxyl. Suitable inorganic bases for this alkylation include potassiumcarbonate and sodium carbonate. Suitable solvents for this alkylationare non-reactive polar organic solvents such as methyl ethyl ketone anddimethyl-formamide. The ester is hydrolyzed using standard syntheticmethods, such as by reaction of the alkylated intermediate with anaqueous acid or base. For example, the ethyl ester is readily hydrolyzedby reaction with 5N sodium hydroxide in a water miscible organicsolvent, such as methanol. Acidification of the reaction withconcentrated hydrochloric acid produces the Formula III compound,wherein R⁶ is hydroxyl, as the hydrochloride salt.

The Formula III compounds, wherein R⁶ is chloro or bromo, are preparedby halogenating the Formula III compounds wherein R⁶ is hydroxyl.Suitable halogenating agents include oxalyl chloride, thionyl chloride,thionyl bromide, phosphorous tribromide, triphosgene, and phosgene.Preferably, R⁶ is chloro. Suitable solvents for this reaction includemethylene chloride, 1,2-dichlorobenzene, chlorobenzene, and1,2-dichloroethane. Preferably, the halogenation reaction is carried outin the same solvent as the subsequent acylation reaction. A catalyticamount of dimethylformamide, from about 0.05 to about 0.25 equivalents,is added to the chlorination reaction. When the reaction is carried outin 1,2-dichloroethane, the reaction is complete after about 2 to 5 hoursat about 47° C. The Formula III compounds, wherein R⁶ is chloro, may bestored as a solid, or as a solution or mixture in methylene chloride,chlorobenzene, 1,2-dichlorobenzene, or 1,2-dichloroethane. Preferably,the chlorination reaction and acylation reaction are carried outsuccessively in the same reaction vessel.

The 2-aryl-6-hydroxy-3-(4-(2-aminoethoxy)benzoyl[b]-thiophenes can beprepared by acylation and subsequent dealkylation of the phenolic groupsin two distinct steps, or sequentially in a “one-pot” reaction. Thestep-wise synthesis is described in the following paragraphs.

The acylated benzothiophene intermediate, a Formula IV compound, isprepared as shown in Scheme III, wherein R⁴, R⁵, R⁶, and HX are asdefined above.

Generally, benzothiophene intermediate II is acylated with a Formula IIIcompound, using boron trichloride or boron tribromide as the acylationcatalyst. The reaction is carried out in an organic solvent, such aschlorobenzene, methylene chloride, 1,2-dichloroethane,1,2-dichlorobenzene, bromobenzene, chloroform,1,1,2,2-tetrachloro-ethane, 1,2,3-trichloropropane, and fluorobenzene.Preferably, the acylation is carried out in methylene chloride,chloro-benzene, or 1,2-dichloroethane. Most preferably, the acylationstep is carried out in methylene chloride. The rate of acylation of theFormula II compound and the rate of dealkylation of the phenolic ethersof the Formula II and IV compounds varies with the choice of solvent,temperature of reaction, and choice of boron trihalide. Because theFormula II compounds having one or more unprotected phenolic groups willnot acylate readily under these conditions, the amount of dealkylationmust be minimized. Because boron tribromide is more preferred fordealkylation of phenolic ethers, the preferred boron trihalide forcatalyzing acylation is boron trichloride. For borontrichloride-catalyzed reactions in methylene chloride, the acylationreaction can be performed at room temperature, with minimal dealkylationof the Formula II and IV compounds. In other solvents, the acylationreaction is carried out at lower temperatures, such as −10° C. to 10°C., to minimize the amount of dealkylation of the reaction startingmaterial and product. When R⁶ is chloro, at least 2 molar equivalents ofthe boron trihalide reagent are required for acylation. When the benzoicacid is used as an acylating agent (R⁶═OH), five equivalents of theboron trihalide are typically used. The Formula IV compound may beisolated as the hydrochloride or hydrobromide salt, or as the free base.

In the step-wise process, the acylated intermediate (Formula IVcompound) is dealkylated to produce the Formula I compound as shown inScheme IV, wherein R⁴, R⁵, and HX are as defined above.

The Formula I compound is produced by reacting the hydrochloride orhydrobromide salt of the Formula IV compound with boron tribromide orboron trichloride. The preferred boron trihalide for dealkylation isboron tribromide. This dealkylation reaction can be carried out in avariety of organic solvents, such as methylene chloride, chlorobenzene,1,2-dichloroethane, chloroform, 1,1,2,2-tetrachloroethane,1,2,3-trichloropropane, 1,2-dichlorobenzene, and fluorobenzene. Thepreferred solvent is 1,2-dichloroethane. When the acid addition salt isused as a starting material, the amount of by-product resulting fromdealkylation of the aminoethyl group is minimized. When methylenechloride is used as the solvent and the boron reagent is borontrichloride, the reaction is generally carried out at a temperature ofabout 55° C. to about 75° C., producing the Formula I compound with nodetectable cleavage of the aminoethyl group. In other solvents, such aschloroform, 1,2-dichloroethane, chlorobenzene, 1,2-dichlorobenzene, andfluorobenzene, the dealkylation occurs readily at ambient temperatures.For example, when 1,2-dichloroethane is the solvent, the reaction isgenerally carried out at 25° C. to 35° C. with no detectable cleavage ofthe aminoethyl group. At least four equivalents of the boron trihalidereagent are typically used for complete reaction within a reasonabletime.

Preferably, the Formula I compounds are prepared by a “one-pot”synthesis from the Formula II and III compounds as shown in Scheme V,wherein R⁴, R⁵, R⁶, and HX are as defined above.

The benzothiophene Formula II compound is acylated with the Formula IIIcompound in the presence of boron trichloride or boron tribromide; borontrichloride is preferred for the “one-pot” process. The reaction can becarried out in a variety of organic solvents, such as chloroform,methylene chloride, 1,2-dichloroethane, 1,2,3-dichloropropane,1,1,2,2-tetrachloroethane, 1,2-dichloro-benzene, chlorobenzene, andfluorobenzene. The preferred solvent for this synthesis is1,2-dichloroethane. The reaction is carried out at a temperature ofabout −10° C. to about 25° C., preferably at 0° C. The reaction is bestcarried out at a concentration of the benzothiophene Formula II compoundof about 0.2 M to about 1.0 M. The acylation reaction is generallycomplete after about two hours to about eight hours.

The acylated benzothiophene, the Formula IV compound, is converted to aFormula I compound without isolation. This conversion is performed byadding additional boron trihalide and heating the reaction mixture.Preferably, two to five molar equivalents of boron trichloride are addedto the reaction mixture, most preferably three molar equivalents. Thisreaction is carried out at a temperature of about 25° C. to about 40°C., preferably at 35° C. The reaction is generally complete after about4 to 48 hours. The acylation/dealkylation reaction is quenched with analcohol or a mixture of alcohols. Suitable alcohols for use in quenchingthe reaction include methanol, ethanol, and isopropanol. Preferably, theacylation/dealkylation reaction mixture is added to a 95:5 mixture ofethanol and methanol (3A). The 3A ethanol can be at room temperature orheated to reflux, preferably at reflux. When the quench is performed inthis manner, the Formula I compound conveniently crystallizes from theresulting alcoholic mixture. Generally, 1.25-3.75 mL of alcohol permillimole of the benzothiophene starting material are used.

The preparation of a solvate of the Formula I compound, wherein HX isHCl, was described previously. Jones et al., J. Med. Chem., 27, 1057(1984). The crystalline product of this “one-pot” process, when BCl₃ isused, is isolated as the solvate of the hydrochloride salt. Thesecrystalline solvates are obtained under a variety of conditions.Generally, the form of the product of the present process is determinedby choice of acylation/dealkylation solvent, boron trihalide, andwork-up conditions.

A particularly useful solvate of the formula compound is the1,2-dichloroethane solvate. This solvate is prepared by carrying out the“one-pot” acylation/dealkylation process in 1,2-dichloroethane. When HXis HCl, the 1,2-dichloroethane solvate of a compound of formula I canexist in two distinct forms. One crystalline solvate form, termedcrystal form I, is prepared by quenching the boron trichloride-catalyzedacylation/dealkylation reaction with ethanol. Preferably, a mixture ofethanol and methanol (95:5) is used in the preparation of this crystalform. This particular crystal form is characterized by the X-raydiffraction pattern shown in Table 1.

TABLE 1 X-ray Diffraction Pattern for Crystal Form 1. d-line spacingI/I_(o) (Angstroms) (×100) 16.1265 3.80 10.3744 8.63 8.3746 5.29 7.988336.71 7.2701 5.06 6.5567 70.77 6.2531 6.79 5.5616 24.05 5.3879 100.005.0471 89.64 4.7391 85.96 4.6777 39.36 4.6332 62.60 4.5191 77.56 4.286736.82 4.2365 41.66 4.1816 49.60 4.0900 11.28 3.9496 11.85 3.7869 36.253.7577 56.16 3.6509 40.62 3.5751 15.65 3.5181 21.52 3.4964 18.53 3.436133.60 3.3610 6.21 3.3115 4.95 3.2564 7.36 3.2002 3.80 3.1199 15.773.0347 14.84 2.8744 9.67 2.8174 10.82 2.7363 11.51

The amount of6-hydroxy-2-(4-hydroxyphenyl)-3-[4-(2-piperidinoethoxy)benzoyl]benzo[b]thiophenehydrochloride present in the crystalline material is about 87.1%, asdetermined using the high performance liquid chromatography (HPLC) assaydescribed below. The amount of 1,2-dichloroethane present in thecrystalline material is about 0.55 molar equivalents, as determined byproton nuclear magnetic resonance spectroscopy.

A large, analytically pure single crystal of the form I1,2-dichloroethane solvate was prepared for single crystal X-rayanalysis. This single crystal was prepared by placing a saturatedmethanolic solution of6-hydroxy-2-(4-hydroxyphenyl)-3-[4-(2-piperidinoethoxy)benzoyl]benzo[b]thiophene hydrochloride in an atmosphere saturated with1,2-dichloroethane (see Example 8). A total of 8419 reflections with 20less than 116° were collected, and used to solve the structure. TheX-ray structure clearly shows that the crystalline material is a1,2-dichloroethane solvate having a 1:2 ratio of solvent to solutemolecules. The theoretical X-ray powder diffraction pattern spectrum,calculated from the single crystal X-ray data, is identical to thatlisted in Table 1, indicating that both solvates are identical.

A second crystalline solvate form, termed crystal form II, is similar tocrystal form I. This second form is prepared by quenching the borontrichloride-catalyzed acylation/dealkylation reaction carried out in1,2-dichloroethane with methanol. Alternatively, the borontrichloride-catalyzed acylation/dealkylation reaction using1,2,3-trichloropropane as the solvent, produces a 1,2,3-trichloropropanesolvate of this form. This particular crystal form is characterized bythe X-ray diffraction pattern shown in Table 2.

TABLE 2 X-ray Diffraction Pattern for Crystal Form II. d-line spacingI/I_(o) (Angstroms) (×100) 10.4311 22.64 8.9173 10.73 8.4765 5.31 8.009550.39 7.3068 4.23 6.6094 79.23 5.6196 22.34 5.4223 89.86 5.1959 11.815.0746 74.90 4.8017 100.00 4.7262 57.97 4.6569 53.35 4.5378 96.75 4.437610.83 4.3397 56.89 4.2782 48.23 4.2129 40.94 4.1037 12.80 3.9880 14.763.8863 8.17 3.7999 42.13 3.7662 57.09 3.6738 38.58 3.5701 18.50 3.539319.00 3.4622 39.57 3.3867 5.02 3.3321 4.33 3.2686 6.79 3.1535 14.863.0450 13.58 2.9028 12.30 2.8302 19.59 2.7544 12.30 2.6366 6.89

The amount of6-hydroxy-2-(4-hydroxyphenyl)-3-[4-(2-piperidinoethoxy)benzoyl]benzo[b]thiophenehydrochloride present in the crystalline material is about 86.8%. Theamount of 1,2-dichloroethane present in the crystalline material isabout 6.5%, as determined by gas chromatography.

The formula I compounds form a variety of distinct solvates witharomatic solvents. A useful aromatic solvate of this compound is thechlorobenzene solvate, which exists in a distinct form termed crystalform III. This particular crystal form is characterized by the X-raydiffraction pattern shown in Table 3.

TABLE 3 X-ray Diffraction Pattern for Crystal Form III. d-line spacingI/I_(o) (Angstroms) (×100) 14.3518 7.24 10.3335 6.17 8.8305 4.29 7.947538.16 6.5904 64.25 6.2848 6.52 5.6048 28.06 5.4107 100.00 5.1544 11.265.0493 53.26 5.0224 46.11 4.8330 76.94 4.7694 34.23 4.6461 50.22 4.575438.61 4.4953 72.65 4.3531 49.15 4.2940 41.64 4.2425 35.75 4.1856 21.634.1338 9.47 4.0793 12.69 3.9960 18.50 3.9037 9.03 3.7854 40.39 3.752154.16 3.6787 28.60 3.6509 17.96 3.5444 31.72 3.4679 41.55 3.3899 7.693.3101 5.72 3.2561 7.42 3.1784 15.19 3.0445 11.17 3.0146 8.94 2.916011.89 2.8217 18.23 2.7500 12.06 2.6436 9.65 2.6156 6.97

The amount of6-hydroxy-2-(4-hydroxyphenyl)-3-[4-(2-piperidinoethoxy)benzoyl]benzo[b]thiophenehydrochloride present in the crystalline material is about 78.6%. Theamount of chlorobenzene present in the crystalline material is about12.3%, as determined by HPLC.

A fourth crystalline solvated form is termed crystal form IV. Thisparticular form is prepared by the boron trichloride-catalyzedacylation/dealkylation process using methylene chloride or chloroform asthe solvent. This particular crystal form is characterized by the X-raydiffraction pattern shown in Table 4.

TABLE 4 X-ray Diffraction Pattern for Crystal Form IV. d-line spacingI/I_(o) (Angstroms) (×100) 10.3696 14.40 8.9032 10.19 8.3125 7.61 7.981841.03 7.2036 7.34 6.5411 74.18 6.2367 6.39 5.5539 20.11 5.3689 100.005.0272 95.92 4.7085 89.13 4.6406 73.37 4.6199 77.58 4.5347 69.70 4.481849.86 4.2589 47.69 4.2067 44.43 4.1659 44.16 4.0957 11.96 3.9347 11.283.7818 40.90 3.7614 53.53 3.6375 36.68 3.5773 20.11 3.5037 25.14 3.440932.34 3.4270 39.54 3.3088 12.64 3.2611 9.65 3.1046 12.77 3.0263 17.532.8536 8.29 2.8131 12.09 2.7309 8.97

The amount of6-hydroxy-2-(4-hydroxyphenyl)-3-[4-(2-piperidinoethoxy)benzoyl]benzo[b]thiophenehydrochloride present in the crystalline material is about 80.4%, asdetermined by HPLC analysis. The amount of chloroform present in thecrystalline material is about 0.42 molar equivalents, as determined byproton nuclear magnetic resonance spectroscopy.

A preferred crystalline form of6-hydroxy-2-(4-hydroxyphenyl)-3-[4-(2-piperidinoethoxy)benzoyl]benzo[b]-thiophenehydrochloride is a non-solvated crystal form. This particular form ispreferred for use in the preparation of pharmaceutical formulationsbecause of the absence of solvent that could affect the patient. Thisparticular crystal form is prepared by recrystallization of the solvatedhydrochloride salt produced by the boron trichloride-catalyzedacylation/dealkylation process. Generally, the solvated hydrochloridesalt is added to a solution of sodium hydroxide in methanol or a mixtureof methanol and water. At least one equivalent of base is used fordissolution and to ensure that the hydrochloride salt is converted tothe free base. Activated carbon is optionally added to the resultingsolution to facilitate removal of impurities. The mixture is filtered toremove the activated carbon, if present, and any insoluble impurities.The filtrate is extracted with an aliphatic hydrocarbon solvent, such ashexane or heptane, to remove the organic solvent used in theacylation/dealkylation reaction. The methanol solution is acidified withhydrochloric acid, such as gaseous or aqueous hydrochloric acid, causingcrystallization of6-hydroxy-2-(4-hydroxyphenyl)-3-[4-(2-piperidinoethoxy)benzoyl]benzo[b]-thiopheneas the non-solvated hydrochloride salt. The resulting crystalline slurryis preferably stirred at ambient temperature for about one to about twohours to ensure complete crystallization. The non-solvated crystallineform is isolated by filtration, followed by drying in vacuo. Thisparticular crystal form is characterized by the X-ray diffractionpattern shown in Table 5.

TABLE 5 X-ray Diffraction Pattern for Non-solvated Crystal Form. d-linespacing I/I_(o) (Angstroms) (×100) 13.3864 71.31 9.3598 33.16 8.46252.08 7.3888 7.57 6.9907 5.80 6.6346 51.04 6.1717 29.57 5.9975 5.675.9135 9.87 5.6467 38.47 5.4773 10.54 5.2994 4.74 4.8680 4.03 4.79105.98 4.6614 57.50 4.5052 5.75 4.3701 9.03 4.2516 69.99 4.2059 57.644.1740 65.07 4.0819 12.44 3.9673 22.53 3.9318 100.00 3.8775 9.07 3.709633.38 3.6561 21.65 3.5576 3.36 3.5037 7.97 3.4522 18.02 3.4138 4.653.2738 10.23 3.1857 8.90 3.1333 6.24 3.0831 9.43 3.0025 12.13 2.94374.96 2.8642 7.70 2.7904 11.95 2.7246 3.05 2.6652 3.32 2.5882 7.30

The amount of6-hydroxy-2-(4-hydroxyphenyl)-3-[4-(2-piperidinoethoxy)benzoyl]benzo[b]thiophenehydrochloride present in the crystalline material is at least 95%.

This non-solvated crystalline material is more pure than the materialproduced by the processes described in the above-referenced patents.This material is free of aluminum impurities, as well as, chlorinatedaliphatic hydrocarbon solvents and aromatic solvents. This non-solvatedcrystalline form is particularly preferred for use in the manufacture ofpharmaceutical compositions.

The following examples further illustrate the processes described. Theexamples are not intended to be limiting to the scope of the inventionin any respect, and should not be so construed. All experiments were rununder positive pressure of dry nitrogen. All solvents and reagents wereused as obtained. The percentages are generally calculated on a weight(w/w) basis; except for HPLC solvents which are calculated on a volume(v/v) basis. Proton nuclear magnetic resonance (¹H NMR) spectra wereobtained on a Bruker AC-300 FTNMR spectrometer at 300.135 MHz. Meltingpoints were determined by differential scanning calorimetry (DSC) in aTA Instrument DCS 2920 using a closed cell and a heating rate of 2°C./minute. The X-ray powder diffraction spectra were obtained in aSiemens D5000 X-Ray Powder Diffraktometer, using copper radiation and aSi(Li) detector.

The reactions were generally monitored for completion using highperformance liquid chromatography (HPLC). The reaction producing theacid chloride, the Formula III compound wherein R⁶ is chloro, wasmonitored using a Zorbax Rx-C8 column, (25 cm×4.6 mm ID, 5μ particle)eluting with a mixture of 60 mM phosphate (KH₂PO₄) and 10 mMoctanesulfonate (pH 2.0)/acetonitrile (60:40). The Formula III compoundwas derivatized with aniline, and analyzed using a carbanilide referencestandard, derived from reaction of phosgene with aniline. A carbanilidestandard stock solution was prepared by dissolving carbanilide (10 mg)and aniline (3 mL). This solution was diluted to a volume of 100 mL withthe eluent described above. The reaction was monitored by the additionof about 0.3 mL of the acid chloride solution to 1 mL of HPLC grademethanol. The resulting mixture was shaken vigorously and allowed toderivatize. After 30 minutes, acetonitrile (6 mL) was added followed bydilution to 100 mL with the eluent described above.

The acylation, dealkylation, or acylation/dealkylation reactions arealso monitored for completion by HPLC. A sample of the reaction mixturewas assayed using a Zorbax Rx-C8 column, (25 cm×4.6 mm ID, 5μ particle),eluting with a gradient as shown below:

Gradient Solvent System Time (min.) A (%) B (%) 0 60 40 5 60 40 10 45 5520 38 62 25 45 55 32 45 55 37 60 40 42 60 40 A: 0.05 M HClO₄ (pH = 2.0)B: acetonitrile

The reaction mixture was analyzed by diluting a 0.1 to 0.2 mL sample to50 mL with a 60:40 mixture of A/B. Similarly, the mother liquor of therecrystallizations was sampled in a similar manner.

The amount (percentages) of6-hydroxy-2-(4-hydroxyphenyl)-3-[4-(2-piperidinoethoxy)benzoyl]benzo[b]thiophenehydrochloride in the crystalline material (potency) was determined bythe following method. A sample of the crystalline solid (5 mg) wasweighed into a 100-mL volumetric flask, and dissolved in a 70/30 (v/v)mixture of 75 mM potassium phosphate buffer (pH 2.0) and acetonitrile.An aliquot of this solution (10 μL) was assayed by high performanceliquid chromatography, using a Zorbax Rx-C8 column (25 cm×4.6 mm ID, 5μparticle) and UV detection (280 nm). The following gradient solventsystem is used:

Gradient Solvent System (Potency) Time (min) A (%) B (%) 0 70 30 12 7030 14 25 75 16 70 30 25 70 30 A: 75 mM KH₂PO₄ buffer (pH 2.0) B:acetonitrile

The percentage of6-hydroxy-2-(4-hydroxyphenyl)-3-[4-(2-piperidinoethoxy)benzoyl]benzo[b]thiophenehydrochloride in the sample is calculated using the peak area, slope(m), and intercept (b) of the calibration curve with the followingequation:

${\% \mspace{14mu} {potency}} = {\frac{{{peak}\mspace{14mu} {area}} - b}{m} \times \frac{{sample}\mspace{14mu} {volume}\mspace{14mu} ({mL})}{{sample}\mspace{14mu} {weight}\mspace{14mu} ({mg})}}$

The amount (percentage) of solvent, such as methanol, ethanol, or1,2-dichloroethane, present in the crystalline material is determined bygas chromatography. A sample of the crystalline solid (50 mg) wasweighed into a 10-mL volumetric flask, and dissolved in a solution of2-butanol (0.025 mg/mL) in dimethylsulfoxide. A sample of this solutionwas analyzed on a gas chromatograph using a DB Wax column (30 m×0.53 mmID,

1μ particle), with a column flow of 10 mL/min and flame ionizationdetection. The column temperature was heated from35° C. to 230° C. over a 12 minute period. The amount of solvent wasdetermined by comparison to the internal standard (2-butanol), using thefollowing formula:

${\% \mspace{14mu} {solvent}} = {\frac{C}{D} \times \frac{E}{F} \times \frac{G}{H} \times I}$

wherein:

C=ratio of solvent in sample

D=average ratio of standard for specific solvent

E=average weight of standard

F=weight of sample (mg)

G=volume of sample (10 mL)

H=volume of standard (10,000 mL)

I=purity of standard (%)

Preparation 1 6-Methoxy-2-(4-methoxyphenyl)benzo[b]thiophene

A solution of 3-methoxybenzenethiol (100 grams) and potassium hydroxide(39.1 grams) in water (300 mL) was added to denatured ethanol (750 mL),and the resulting mixture cooled to about 0° C. The cold mixture wastreated with 4′-methoxyphenacyl bromide (164 grams) in several smallportions. Upon complete addition, the mixture was cooled for anadditional ten minutes, then allowed to warm to room temperature. Afterthree hours, the mixture was concentrated in vacuo, and the residuetreated with water (200 mL). The resulting mixture was treated withethyl acetate, and the phases were separated. The organic phase waswashed with water (2×), sodium bicarbonate solution (2×), and sodiumchloride solution (2×). The organic phase was then dried over magnesiumsulfate, filtered, and evaporated to dryness in vacuo to give 202 gramsof a-(3-methoxyphenylthio)-4-methoxyacetophenone. This crude product wascrystallized from methanol and washed with hexane to give 158 grams.Melting point 53° C.

Polyphosphoric acid (930 grams) was heated to 85° C. and treated withthe intermediate product from above (124 grams) in small portions over30 minutes. Upon complete addition, the resulting mixture was stirred at90° C. After an additional 45 minutes, the reaction mixture was allowedto cool to room temperature. This mixture was treated with crushed icewhile the mixture was cooled in an ice bath. The resulting mixture wastreated with water (100 mL) producing a light pink precipitate. Theprecipitate was isolated by filtration, washed with water and methanol,and dried in vacuo at 40° C. to give 119 grams of6-methoxy-2-(4-methoxyphenyl)benzo[b]thiophene. This crude product wasslurried in hot methanol, filtered, and washed with cold methanol. Theresulting solid material was recrystallized from ethyl acetate (4liters), filtered, washed with hexane, and dried in vacuo to 68 grams ofthe title compound. Melting point 187-190.5° C.

Preparation 2 Ethyl 4-(2-Piperidinoethoxy)benzoate

A mixture of ethyl 4-hydroxybenzoate (8.31 g),1-(2-chloroethyl)piperidine monohydrochloride (10.13 g), potassiumcarbonate (16.59 g), and methyl ethyl ketone (60 ml) was heated to 80°C. After one hour, the mixture was cooled to about 55° C. and treatedwith additional 1-(2-chloroethyl)piperidine mono-hydrochloride (0.92 g).The resulting mixture was heated to 80° C. The reaction was monitored bythin layer chromatography (TLC), using silica-gel plates and ethylacetate/acetonitrile/triethylamine (10:6:1, v/v). Additional portions of1-(2-chloroethyl)piperidine hydrochloride are added until the starting4-hydroxybenzoate ester is consumed. Upon complete reaction, thereaction mixture was treated with water (60 mL) and allowed to cool toroom temperature. The aqueous layer was discarded and the organic layerconcentrated in vacuo at 40° C. and 40 mm Hg. The resulting oil was usedin the next step without further purification.

Preparation 3 4-(2-Piperidinoethoxy)benzoic Acid Hydrochloride

A solution of the compound prepared as described in Preparation 2 (about13.87 g) in methanol (30 mL) was treated with 5 N sodium hydroxide (15mL), and heated to 40° C. After

4½ hours, water (40 mL) was added. The resulting mixture was cooled to5-10° C., and concentrated hydrochloric acid (18 mL) was added slowly.The title compound crystallized during acidification. This crystallineproduct was collected by filtration, and dried in vacuo at 40-50° C. togive 83% yield of the title compound. Melting point 270-271° C.

Preparation 4 4-(2-Piperidinoethoxy)benzoyl Chloride Hydrochloride

A solution of the compound prepared as described in Preparation 3 (30.01g) and dimethylformamide (2 mL) in methylene chloride (500 mL) wastreated with oxalyl chloride (10.5 mL) over a 30-35 minute period. Afterstirring for about 18 hours, the reaction was assayed for completion byHPLC analysis. Additional oxalyl chloride may be added to the reactionif the starting carboxylic acid is present. Upon completion, thereaction solution was evaporated to dryness in vacuo. The residue wasdissolved in methylene chloride (200 mL), and the resulting solutionevaporated to dryness. This dissolution/evaporation procedure wasrepeated to give the title compound as a solid. The title compound maybe stored as a solid or as a 0.2 M solution in methylene chloride (500mL).

Example 16-Methoxy-2-(4-methoxyphenyl)-3-[4-(2-piperidinoethoxy)benzoyl]-benzo[b]thiopheneHydrochloride

A mixture of the compound prepared as described in Preparation 1 (8.46grams) and the acid chloride prepared as described in Preparation 3(10.0 grams) in methylene chloride (350 mL) was cooled to about 20-25°C. The cool mixture was treated with boron trichloride (2.6 mL), and theresulting mixture mechanically stirred. The reaction was monitored byHPLC using the assay described above. After 85 minutes, the in situ HPLCyield based on a6-methoxy-2-(4-methoxyphenyl)-3-[4-(2-piperidinoethoxy)benzoyl]benzo[b]thiophenestandard was 88%.

Example 26-Hydroxy-2-(4-hydroxyphenyl)-3-[4-(2-piperidinoethoxy)benzoyl]-benzo[b]thiopheneHydrochloride 1,2-Dichloroethane Solvate (Crystal Form I)

A solution of6-methoxy-2-(4-methoxyphenyl)-3-[4-(2-piperidinoethoxy)benzoyl]benzo[b]thiophenehydrochloride (2.0 g) in 1,2-dichloroethane (20 mL) was treated withboron trichloride (2.0 mL). The resulting mixture was stirred at 35° C.for about 18 hours. A mixture of ethanol and methanol (10 mL, 95:5, 3A)was treated with the reaction mixture from above, causing the alcoholicmixture to reflux. Upon complete addition, the resulting crystallineslurry was stirred at 25° C. After one hour, the crystalline product wasfiltered, washed with cold ethanol (10 mL), and dried at 40° C. in vacuoto give 1.78 g of the title compound. The X-ray powder diffractionpattern is identical to that reported in Table 1.

Potency: 80.2%

1,2-Dichloroethane: 7.5% (gas chromatography)

Example 36-Hydroxy-2-(4-hydroxyphenyl)-3-[4-(2-piperidinoethoxy)benzoyl]-benzo[b]thiopheneHydrochloride Methylene Chloride Solvate (Crystal Form IV)

A mixture of the compound prepared as described in Preparation 1 (7.54grams) in methylene chloride (10 mL) and the acid chloride prepared asdescribed in Preparation 4 (140 mL, 0.21 M solution in methylenechloride) was placed in a sealed reaction vessel (Hastalloy Parr). Thesolution was cooled to 0° C. and treated with boron trichloride (7.2mL). The resulting reaction mixture was stirred at room temperature.After three hours, the reaction was cooled in an ice bath for 10minutes. A second portion of boron trichloride (4.8 mL) was added to thereaction mixture, and the mixture was heated to 75° C. After 2.5 hours,the reaction mixture was cooled to about 15° C. The cool mixture wastreated with tetrahydrofuran (15 mL) and methanol (45 mL). This mixturewas stirred for about one hour at 18° C., producing a crystalline solid.The crystalline solid was removed by filtration, rinsed with coldmethanol (45 mL), and dried in vacuo at 40° C. for 18 hours, to give12.5 grams of the title compound. The X-ray powder diffraction patternis identical to that reported in Table 4. Melting point 207° C.

Potency: 81.8%

Methylene chloride: 0.4 molar equivalents (¹H NMR)

Example 46-Hydroxy-2-(4-hydroxyphenyl)-3-[4-(2-piperidinoethoxy)benzoyl]-benzo[b]thiopheneHydrochloride 1,2-Dichloroethane Solvate (Crystal Form I)

A mixture of the compound prepared as described in Preparation 2 (15 g)and dimethylformamide (0.2 mL) in 1,2-dichloroethane (250 mL) was cooledto 0° C. Phosgene (8.25 mL) was condensed in a cold, jacketed additionfunnel (−10° C.), and added to the cold mixture over a period of twominutes. The resulting mixture was heated to about 47° C. After abouttwo and one half hours, the reaction was assayed by HPLC for completion.Additional phosgene may be added to drive the reaction to completion.Excess phosgene was removed by vacuum distillation at 30-32° C. and105-110 mm Hg.

After about three to four hours, the reaction solution was treated withthe compound prepared as described in Preparation 1 (13.52 g). Theresulting solution was cooled to 0° C. Boron trichloride (12.8 mL) wascondensed in a graduated cylinder, and added to the cold reactionmixture. After eight hours at 0° C., the reaction solution was treatedwith additional boron trichloride (12.8 mL). The resulting solution washeated to 30° C. After 15 hours, the reaction was monitored forcompletion by HPLC.

A mixture of ethanol and methanol (125 mL, 95:5, 3A) was heated toreflux, and treated with the reaction solution from above over a 60minute period. Upon complete addition, the acylation/demethylationreaction flask was rinsed with additional ethanol (30 mL). The resultingslurry was allowed to cool to room temperature with stirring. After onehour at room temperature, the crystalline product was filtered, washedwith ethanol (75 mL), and dried at 40° C. in vacuo to give 25.9 g of thetitle compound. The X-ray powder diffraction pattern is reported inTable 1. Melting point 261° C.

Potency: 87.1%

1,2-Dichloroethane: 0.55 molar equivalents (¹H NMR)

Example 56-Hydroxy-2-(4-hydroxyphenyl)-3-[4-(2-piperidinoethoxy)benzoyl]-benzo[b]thiopheneHydrochloride Chlorobenzene Solvate (Crystal Form 3)

A solution of the compound prepared as described in Preparation 1 (2.92grams) and the acid chloride prepared as described in Preparation 4(3.45 grams) in chlorobenzene (52 mL) was cooled to about 0° C. The coldsolution was treated with boron trichloride (2.8 mL). The resultingmixture was mechanically stirred at about 0° C. After three hours,additional boron trichloride (2.8 mL) was added, and the reactionmixture was allowed to warm to room temperature. After about 16-20hours, the reaction mixture was cooled to 0° C. The cold reaction wasquenched by the slow addition of ethanol (26 mL) over 30 minutes. Duringthe addition of the alcohol, a crystalline solid formed. Upon completeaddition of the alcohol, the resulting mixture was stirred at roomtemperature for one hour. The crystalline solid was removed byfiltration, washed with cold ethanol (25 mL), and dried in vacuo at 40°C. to give 5.94 grams of the title compound as a yellow solid. The X-raypowder diffraction pattern is identical to that reported in Table 3.Melting point 247° C.

Potency: 78.6%

Chlorobenzene: 12.3% (HPLC)

Example 66-Hydroxy-2-(4-hydroxyphenyl)-3-[4-(2-piperidinoethoxy)benzoyl]-benzo[b]thiopheneHydrochloride 1,2-Dichloroethane Solvate (Crystal Form II)

A mixture of the compound prepared as described in Preparation 1 (2.92g), the compound prepared as described in Preparation 4 (3.45 g), and1,2-dichloroethane (52 mL) was cooled to about 0° C. Boron trichloridegas was condensed into a cold graduated cylinder (2.8 mL), and added tothe cold mixture described above. After eight hours at 0° C., thereaction mixture was treated with additional boron trichloride (2.8 mL).The resulting solution was heated to 35° C. After 16 hours, the reactionwas complete.

Methanol (30 mL) was treated with the reaction mixture from above over a20-minute period, causing the methanol to reflux. The resulting slurrywas stirred at 25° C. After one hour, the crystalline product wasfiltered, washed with cold methanol (8 mL), and dried at 40° C. in vacuoto give 5.14 g of the title compound. The X-ray powder diffractionpattern is reported in Table 2.

Melting point 225° C.

Potency: 86.8%

1,2-Dichloroethane: 6.5% (gas chromatography)

Example 76-Hydroxy-2-(4-hydroxyphenyl)-3-[4-(2-piperidinoethoxy)benzoyl]-benzo[b]thiopheneHydrochloride

The compound prepared as described in Example 4 (4.0 grams) andactivated carbon (0.4 grams, Darco G-60, Aldrich Chem. Co., Inc.,Milwaukee, Wis.) were slurried in methanol (50 mL) at room temperature.The resulting mixture was treated with a solution of sodium hydroxide(0.313 grams) in methanol (10 mL). After 30 minutes, the slurry wasfiltered through Whatman #1 filter paper precoated with diatomaceonsearth (Hyflo Super Cel®, Aldrich Chem. Co.). The filter cake was rinsedwith methanol (5 mL). The combined filtrate was treated (dropwise) with2N hydrochloric acid (4 mL). The resulting slurry was stirred for 30minutes at room temperature, then cooled to about 9° C. After one hour,the cool mixture was filtered. The filter cake was rinsed with coldmethanol (10 mL, OC), and dried in vacuo at 60° C. for about 18 hours togive 2.06 grams of an off-white free flowing powder.

Potency: 96.8%

Related substances: 1.41%

Example 86-Hydroxy-2-(4-hydroxyphenyl)-3-[4-(2-piperidinoethoxy)-benzoyl]benzo[b]thiopheneHydrochloride 1,2-Dichloroethane Solvate (Crystal Form II)

A saturated solution of6-hydroxy-2-(4-hydroxyphenyl)-3-[4-(2-piperidinoethoxy)benzoyl]benzo[b]thiophenehydrochloride was produced by stirring a slurry of the compound preparedas described in Example 7 in methanol at room temperature overnight.This mixture was filtered (Whatman #1 filter paper). A portion of thefiltrate (20-25 mL) was placed in a 50 mL Erlenmeyer flask. This flaskwas placed within a glass jar (3.5 in.×4 in.) containing1,2-dichloroethane (about 10 mL). The jar was sealed and the combinationwas allowed to stand at room temperature. After 24 hours, singlecrystals had crystallized from the methanol solution. These crystalswere filtered and dried in vacuo. The crystal structure was determinedwith a Siemens R3m/V automated four-circle diffractometer usingmonochromatic copper radiation (λ=1.54178 Å). The crystal structure wassolved using the direct methods routine TREF of the SHELXTL PLUS programlibrary. Full-matrix least-squares refinement was conducted withanisotropic temperature factors for all atoms except hydrogens, whichwere included at calculated positions with isotropic temperaturefactors. The final R-factor was 8.02%. The crystal data is shown below.

Crystal Data Space group C2/C Unit all dimensions a = 20.720(7)Å b =9.492(2)Å c = 28.711(4)Å β = 96.50(2)° Volume 5610(2)Å³ Density (calc.)1.409 mg/m³ Absorption coefficient 3.951 mm⁻¹

The X-ray structure clearly shows that the crystalline material is a1,2-dichloroethane solvate having a 1:2 ratio of molecules of1,2-dichloroethane to molecules of6-hydroxy-2-(4-hydroxyphenyl)-3-[4-(2-piperidinoethoxy)benzoyl]-benzo[b]-thiophenehydrochloride.

Example 96-Hydroxy-2-(4-hydroxyphenyl)-3-[4-(2-piperidinoethoxy)-benzoyl]benzo[b]thiopheneHydrochloride 1,2,3-Trichloropropane Solvate (Crystal Form II)

A mixture of the compound prepared as described in Preparation 1 (2.70g), the compound prepared as described in Preparation 4 (3:60 g), and1,2,3-trichloropropane (50 mL) was treated with boron trichloride (2.6mL). After three hours at 20-25° C., the reaction mixture was treatedwith additional boron trichloride (2.6 mL). After about 18 hours, thereaction mixture was treated with tetrahydrofuran (15 mL) followed bythe slow addition of methanol (15 mL). After these additions werecomplete, the resulting mixture was stirred at room temperature. Afterone hour, the crystalline solid was collected by filtration, washed withcold methanol (10 mL), and dried at 50° C. in vacuo to give 4.13 g ofthe title compound. The X-ray powder diffraction pattern was identicalto that reported in Table 2. Melting point 236° C.

Potency: 78.9%

1,2,3-Trichloropropane: 0.5 molar equivalents (1H NMR)

Example 106-Hydroxy-2-(4-hydroxyphenyl)-3-[4-(2-piperidinoethoxy)-benzoyl]benzo[b]thiopheneHydrochloride Chloroform Solvate (Crystal Form IV)

The title compound (4.42 g) was prepared using the procedure describedin Example 9, except the reaction solvent was chloroform (50 mL). TheX-ray powder diffraction pattern was identical to that reported in Table4. Melting point 258° C.

Potency: 80.4%

Chloroform: 0.42 molar equivalents (¹H NMR)

Example 116-Hydroxy-2-(4-hydroxyphenyl)-3-[(4-(2-piperidinoethoxy)-benzoyl]benzo[b]thiopheneHydrochloride

A solution of sodium hydroxide (0.313 g) in methanol (10 mL) was dilutedwith additional methanol (50 mL). This solution was treated with thecompound prepared as described in Example 6 (4.0 g). After 45 minutes atroom temperature, the solution was filtered (Whatman #1 filter paper)and the filter paper rinsed with methanol (3 mL). The filtrate wastreated with 2 N hydrochloric acid (4 mL), producing a crystallineslurry. After 1½ hours, this crystalline product was filtered, washedwith methanol (5 mL), and dried at 45-50° C. in vacuo to give 2.103 g ofthe title compound. The X-ray powder diffraction pattern was identicalto that reported in Table 5. Potency: 96.5%

Example 126-Hydroxy-2-(4-hydroxyphenyl)-3-[4-(2-piperidinoethoxy)-benzoyl]benzo[b]thiopheneHydrochloride

A solution of sodium hydroxide (0.313 g) in methanol (10 mL) was dilutedwith additional methanol (40 mL) and water (10 mL). This solution wastreated with the compound prepared as described in Example 5 (4.0 g).The resulting solution was extracted with hexane (2×50 mL) to remove thechlorobenzene. The methanolic phase was treated with 2 N hydrochloricacid (4 mL), producing a crystalline slurry. After one hour, thecrystalline product was filtered, washed with methanol (5 mL), and driedat 60° in vacuo to give 2.23 g of the title compound. The X-ray powderdiffraction pattern was identical to that reported in Table 5.

Often, compounds which have poor solubility profiles can have theirbioavailability enhanced by increasing the surface area of theformulated particles. The surface area generally increases per unitvolume as the particle size decreases. Various techniques for grindingor milling a drug substance are well known in the art and each of thesetechniques are commonly used to decrease particle size and increase thesurface area of the particle. It would seem reasonable that the best wayto deal with any slightly soluble compound would be to mill it to thesmallest size possible; however, this is not always practical ordesirable. The milling process has an economic cost not only it thedirect cost of the process, itself, but also with other associatedfactors. For example, very finely divided material presents difficultiesand cost in capsule filling or tablet preparation, because the materialwill not flow, but becomes caked in finishing machinery. Such finishingdifficulties generate non-homogeneity in the final product, which is notacceptable for a drug substance. Additionally, the milling process,physically generates heat and pressure on the material, such conditionslead to chemical degradation of the compound, thus such millingtechniques are usually kept to a minimum.

Therefore, there is always dynamic between the properties which yieldthe maximum bioavailability (particle surface area) and the practicallimits of manufacture. The point of compromise which marks this “bestsolution” is unique to each situation and unique as to itsdetermination.

Methods for determining the size of particles are known in the art. Thefollowing is a description of one method, but is not intended to belimiting. For example, the general method of U.S. Pat. No. 4,605,517could be employed.

In preparing the particulate compound of the invention a compound offormula I, in its raw state, is first characterized for size using aninstrument adapted to measure equivalent spherical volume diameter, thatis to say a Horiba LA900 Laser Scattering Particle Size DistributionAnalyzer or equivalent instrument. Typically a representative sample ofa compound of formula I would be expected to comprise in its raw stateparticles having a mean equivalent spherical volume diameter of about110-200 microns and with a broad size distribution.

After being characterized for size in its raw state, the raw compound isthen milled, preferably using a pin mill under suitable conditions ofmill rotation rate and feed rate, to bring the particle size valuewithin the above mentioned limits according to the invention. Theefficiency of the milling is checked by sampling using a Horiba LA900Laser Scattering Particle Size Distribution Analyzer and the finalparticle size is checked in a similar manner. If the first pass throughthe mill does not produce the required size distribution, then one ormore further passes are effected.

The compound of formula I in its particulate form within the abovementioned limits according to the invention may then be mixed with anexcipient or carrier as necessary and, for example, compressed intotablets. Thus, for example, the particulate compound may be mixed withanhydrous lactose, lactose monohydrate, a portion of crospovidone andgranulated in an aqueous dispersion of povidone and polysorbate 80.After drying and milling into granules the material can be terminallyblended with magnesium stearate and remaining crospovidone to becompressed into tablets.

Because the particles in the raw state as well as after milling or otherparticle size reduction techniques are irregular in shape, it isnecessary to characterize them not by measurement of an actual size suchas thickness or length, but by measurement of a property of theparticles which is related to the sample property possessed by atheoretical spherical particle. The particles are thus allocated an“equivalent spherical diameter”.

The values found from characterizing a large number of “unknown”particles can be plotted frequency vs. diameter or in other methodsweight vs. diameter, usually adopting percentage undersize values forfrequency or weight. This gives a characteristic curve representing sizedistribution of the sample, i.e., cumulative percentage undersizedistribution curve. Values from this can be read off directly or plottedon log-probability paper to give an appropriate straight line. The meanequivalent spherical volume diameter is the 50% undersize value.

The mean equivalent spherical volume diameter found is thus astatistical representation of a theoretical particle having the sameproperty as the “unknown” particle.

As indicated above the mean equivalent sphere volume diameter of theparticles of the milled compound of formula I may be evaluated using aHoriba LA900 Laser Scattering Particle Size Distribution Analyzer. Usingsuch an instrument values for a suspension of the particle of unknownsize may be obtained and the instrument may be monitored using a controlsample having particles within the size range expected based onstatistical analysis of the sample. Multiple runs of the control sampleestablished the standard deviation in measurement of the mean to be 1.3microns.

Following is a description by way of example of the preparation ofcompositions in accordance with the invention. In all of the Examplesthe compound was prepared from raw form using a pin mill and consistedof particles having a mean equivalent spherical volume diameter ofbetween about 5 and 20 microns, at least 90% of the particles having aparticle size of less than about 35 microns.

The particle size of the reduced raloxifene HCl was measured as follows.The laser scattering particle size distribution analysis was effected ona small sample of the reduced material which is suspended inapproximately 180 ml of dispersant solution. Sample is added to thedispersant until an acceptable level of laser light obscuration achievedat which point the particle size distribution is measured. Prior to thesample suspension the dispersant solution was prepared by adding 20drops of Coulter 1A dispersant to a saturated aqueous solution ofraloxifene HCl. The dispersant solution was filtered through a 0.2micron microporous membrane filter to provide the necessaryparticle-free suspending dispersant.

Within five minutes of the preparation of the dispersion, triplicateparticle size measurements were performed. Triplicate measurements areeffected as a minimum check a) to produce more reliable measurements andb) to check the equivalent sampling of the suspended material has beenreproducible i.e., the suspension has not settled.

The results were automatically recorded and displayed graphically togive a frequency percentage vs. undersize and a cumulative percentagevs. undersize characteristic curves for the sample. From this, the meanequivalent spherical volume diameter value was derived (50% undersizevalue) together with the standard deviation of the distributioncalculated as above.

Several physical properties of raloxifene hydrochloride have beeninvestigated during the progression of the compound through development.These include particle size, surface area, and powder bulk density.

A primary determinant in the potential influence of such properties ondrug product performance is the aqueous solubility of the drugsubstance. Raloxifene hydrochloride has a water solubility ofapproximately 0.3 mg/mL at 25° C. and significantly lower values inSimulated Gastric Fluid, USP (0.003 mg/mL) and Simulated IntestinalFluid, USP (0.002 mg/mL) at 37*C. The aqueous value falls into the USPclassification of “very slightly soluble”, while according to the recentSUPAC guidance (“Industry Guidance Immediate Release Solid Oral DosageForms Pre- and Post-Approval Changes: Chemistry, Manufacturing andControls, In Vitro Dissolution Testing, and In Vivo BioequivalenceDocumentation”, Prepared by the Immediate Release Scale-Up and PostApproval Change (SUPAC), Expert Working Group of the ChemistryManufacturing Controls Coordinating Committee (CMC CC) of the Center forDrug Evaluation and Research at the FDA) on immediate release solid oraldosage forms, the compound has low solubility with a dose solubilityvolume of greater than 250 mL. Given the low solubility, the rate atwhich the dosage form dissolves in the gastrointestinal tract canpotentially impact the rate and extent of absorption of the activecompound. Two related physical properties of the bulk drug which canalter the dissolution rate of the dosage form are particle size andsurface area. The impact of surface area which is a function of particlesize is illustrated in the Noyes-Whitney equation given below.

dC/dt=(D/h)*(S)*(Cs−C)

Here, C is the concentration of drug at time t, D is the diffusioncoefficient of drug in the medium, h is the thickness of diffusionlayer, Cs is the saturation solubility of drug in the diffusion layerand S is the effective surface area of the drug particles. To ascertainthe effect of particle size/surface area of raloxifene HCl on in vitrodissolution, lots with varying particle size distributions were obtainedvia recrystallization and further modified through various millingtechnologies. The following table contains pertinent data on four bulklots produced in this effort, which includes particle size datagenerated utilizing laser light diffraction, and surface area datacollected by nirogen adsorption, and analyzed through the BET (Brunauer,Emmett, Teller) equation.

TABLE 6 Bulk Milling Surface Area Mean Particle 90% less Lot #Technology m²/gm Size (μm) than (μm) #1 Micronized 6.09 3.9 6.8 #2Recrystallized 2.28 8.4 13.9 #3 Ball Milled 2.10 23.3 55.3 #3 SlurryMilled 0.45 48.1 89

These four bulk lots were handfilled into capsules to provide 60.0 mg ofraloxifene hydrochloride and submitted for dissolution testing in a 0.1%aqueous polysorbate 80 medium utilizing USP Apparatus II, with a paddlespeed of 50 rpm. Data was collected at 10, 20, 30 and 45 minutes toproduce a dissolution profile.

TABLE 7 Time (min.) % Dissolved Time (min.) % Dissolved Lot # 1(micronized) Lot #2 (Control) 10 51 10 41 20 68 20 60 30 78 30 68 45 8845 74 Lot #3 (Ball-milled) Lot #4 (Slurry Milled) 10 31 10 15 20 45 2027 30 54 30 35 45 64 45 49

It was observed that a range of dissolution profiles resulted from thevarious particle size distributions of the bulk drug substance, withvalues ranging from 25% to approximately 80% dissolved at 30 minutes. Inan attempt to evaluate these differences upon in vivo absorption, astudy was conducted in Fischer 344 rats. In the study, rats were dosedwith the same four bulk raloxifene lots in their diet (0.4% w/w) forseven days. Plasma concentrations of unconjugated raloxifene werequantitated by HPLC for the four bulk lots. The following table showsthe excellent linear correlations obtained between the percentraloxifene hydrochloride dissolved at 10 minutes or 30 minutes in the invitro dissolution test to the average area under the curve (AUC,ng-h/mL) values obtained in rats for each of the bulk drug lots.

TABLE 8 % Dissolved at % Dissolved at Lot 10 Minutes 30 Minutes AUC(ng-h/mL) #1 - Micronized 50 78 10056 #2 - As 41 68 8037 Recrystallized#3 - Ball Milled 31 55 5743 #4 - Slurry Milled 15 35 3329

This in vitro to in-vivo correlation supports the discriminating abilityof the dissolution method, as well as emphasizing the need for a controlstrategy for either the particle size distribution or surface area ofthe bulk drug substance. Further evaluation of this data indicated thatthe particle size data correlates better to the differences noted in thedissolution data and in vivo absorption. This can be explained basedupon the Noyes-Whitney equation, which relates dissolution to theeffective surface area. It is postulated that the surface area asmeasured by nitrogen adsorption for the various types of milledraloxifene does not predict the effective surface area accessible to thedissolution medium. This is demonstrated when comparing therecrystallized (control) lot (lot #2) and ball milled lot (lot #3).While they have very similar surface area values, 2.28 and 2.10 m²/gmrespectively, the control lot has a finer mean particle size, 8.4microns compared to 23.3 microns for the ball milled lot. SEMphotomicrographs of the ball milled particles show very irregularsurfaces with numerous cracks and fissures which would result inincreased surface area as measured by nitrogen adsorption, but may notprovide surface area accessible to the dissolution medium, resulting ina lower effective surface area. This reasoning can explain the bettercorrelation of the differences in particle size to the differences in invitro dissolution and in vivo absorption, compared to the similarity ofthe surface areas for the two lots. Based upon these findings, thedecision was made to pursue particle size distribution as a controlparameter to ensure consistent performance of the drug product withregards to release of the drug component.

To further investigate the impact of particle size of raloxifene HCl ondrug product performance as measured by in vitro dissolution and in vivoabsorption, a single dose, plasma concentration versus time study wasdesigned in cynomolgus monkeys. The study compared absorption from twobulk lots of raloxifene which possessed mean particle sizes of 48.1 and9.0 microns. The lots were formulated into granulation matricesrepresentative of granulations being compacted into tablets for humanuse. In addition, the bulk lot with the 9.0 mean particle size wasgenerated through pin milling technology which represents the desiredcommercial milling route. The table below summarizes the particle sizedata of the two bulk lots.

TABLE 9 Bulk Lot/ Mean Granulation Milling 10% Less 50% Less 90% LessParticle Lot # Technology Than μm Than μm Than μm Size Lot 5A Slurry11.4 44.1 90 48.1 Lot 5B Pin 3.2 8.6 15.1 9.0

For the purposes of producing a dissolution profile, the granulationswere handfilled into capsules to provide the equivalent of 60 mg ofraloxifene hydrochloride. The dissolution data produced in 0.1% aqueousTween medium, utilizing the paddle method at 50 rpm, are shown below.

TABLE 10 Time (min.) % Dissolved Lot 5A (slurry milled) 10 33 20 55 3065 45 74 Lot 5B (pin milled) 10 63 20 91 30 95 45 97

These differences in particle size distribution again producedsignificant differences in the dissolution profile in the aqueous 0.1%polysorbate 80 dissolution medium. In the study monkeys received eachformulation according to a crossover study design and incorporating areplicate period to allow for intrasubject variability. The followingTable 11 shows the mean average plasma concentrations of totalraloxifene after the administration of a 25 mg/kg oral dose to themonkeys.

TABLE 11 Lot 5A (slurry milled) Time (hrs.) ng Total Raloxifene/mlplasma  1.4 78  3.6 67  8.2 81 12.1 60 24.3 45 30   32 36.4 22 48.6 14Lot 5B (pin milled) Time (hrs.) ng Raloxifene/ml plasma  1.4 108  3.6 84 8.2 121 12.1 95 24.3 70 30   51 36.4 34 48.6 23

As seen from the plasma concentration versus time profiles given inTable 11, the formulation with the finer particle size bulk drugsubstance provided higher plasma concentrations of total raloxifene atall of the timepoints sampled. The superior absorption from theformulation with the finer particle size is reflected in both the rateand extent of absorption as illustrated in the following summary ofpharmacokinetic parameters from the study.

Lot Number Cmax (ng/mL) AUC (ng-h/mL) 5A (9.0 microns) 131 3608 5B (48.1microns) 96 2357The differences shown were found to be significant upon statisticalanalysis (AUC, p<0.005 and Cmax, p<0.02). This data is further evidenceof the critical nature of the particle size distribution on its impacton bioavailability. The study also confirms the discriminating abilityof the in vitro dissolution method and its relationship to in vivoabsorption. Once again, the differences observed in the in vitrodissolution profiles translated into in vivo absorption differences.

Based upon the above work and physical property data generated, aparticle size specification was established. The invention provides thatthe mean particle size, as determined by laser light diffraction, shouldbe less than about 25 microns. In addition, 90% of the particles byvolume should be under 50 microns, which allows for characterization ofthe distribution. Preferably, the size is between about 5 and about 20microns, and 90% of the particles have a size of less than about 35microns. To justify this range, bulk lots were produced by pin millingand samples of the available extremes were manufactured into formulatedtablets and in vitro dissolution testing. In one study, six bulk lots ofraloxifene hydrochloride (ca. 1 kg) were received and manufactured intoformulated 60 mg raloxifene HCl tablets representative of the tabletsbeing utilized in Phase III clinical testing. The particle size data forthe lots utilized is summarized in the following Table 12.

TABLE 12 (all particle size in microns) 10% Less 50% Less 90% Less Lot #Than Than Than Mean 6 2 6 12 6 7 3 8 21 10 8 3 11 31 15 9 3 12 30 14 102 5 10 6 11 3 9 23 11

The dissolution profile in 0.1% aqueous polysorbate 80 for all of these6 bulk lots formulated into tablets are comparable in all cases. Inaddition, all lots displayed a relatively fast dissolution profile, withvalues greater than 90% dissolved at 20 minutes. To statistically assessthe dissolution as a function of particle size, JMP Statistical andGraphics Guide software (SAS Institute, Inc., Cary, N.C.) was utilizedand a plot was generated where the percent dissolved at 20 minutes wasplotted as a function of the average particle size of each lot. Table 13sets out the data.

TABLE 13 % Dissolved - Raloxifene Hydrochloride from Core Tablets TimeLot Number (Min) 6 7 8 9 10 11 10 89.2 88.1 81.1 74.5 84.1 80.5 20 92.392.6 95.4 91.3 96.0 93.7 30 93.2 93.9 97.0 93.3 96.3 94.0 45 93.0 94.198.4 93.9 96.1 94.6

The scatter observed in the plot, along with the high p-value (0.81)support the conclusion of a non-significant effect of particle size ondissolution over this range of particle sizes. Similar analyses wereperformed at the other timepoint, 10, 30 and 45 minutes, with calculatedp-values of 0.11, 0.76, and 0.40 respectively. These high p-values alongwith the observation of both negative and positive slopes at the varioustimepoints again support the appropriateness of the range for theparticle size.

Another similar study was performed with 7 different particle sizedistributions of bulk drug, with each again being formulated into 60 mgtablets. The particle size data for these lots is summarized in Table14.

TABLE 14 (all particle size in microns) 10% Less 50% Less 90% Less LotThan Than Than Mean 70B 3.3 14.5 39.3 18.8 70E 2.8 10.5 26.3 13.0 70F3.4 16.0 50.2 22.9 71B 3.1 12.9 38.9 17.8 71D 2.8 10.1 25.6 12.6 71G 3.314.6 42.1 19.6 71H 2.9 11.1 28.2 13.7The dissolution data collected in 0.1% aqueous polysorbate 80 for theseseven bulk lots formulated into tablets is given in the following table.

TABLE 15 % Dissolved - Raloxifene Hydrochloride Time Lot Number (Min)70B 70E 70F 71B 71D 71G 71H 10 76 81 73 76 75 61 68 20 94 96 91 93 88 8591 30 98 99 95 98 91 88 95 45 99 99 97 99 97 97 98As with the previous set of particle size distributions, the comparabledissolution profiles obtained with these particle size distributionssupport the ranges for particle size given in this invention.

Given the relationship shown between in vitro dissolution and in vivoabsorption, it follows that the particle size distribution range claimedin this patent will provide surprisingly consistent in vivoabsorption/bioavailability characteristics.

In addition to the role of particle size in vitro dissolution and invivo absorption, another important aspect is its role on the variousunit operations of the drug product manufacturing process. While theparticle size specification ensures consistent delivery of the drugmolecule to the sites of absorption in the gastrointestinal tract, italso allows for better control during the wet granulation step of thetablet manufacturing process. By controlling the particle size, thevariations in quantity of water needed to elicit the appropriateprogression of the granulation power consumption curve is reduced. Bymaintaining the particle size within the previous mentioned constraints,established quantities of water can be dictated in the manufacturingticket for routine lot manufacture. The granulation step is common tomany tablet and capsule manufacturing operations and is typically drivenby the addition of water to bring about the desired endpoint of thegranulation. A downstream unit operation dependent upon the granulationendpoint is the milling of the dried granulation and the resultingparticle size distribution obtained on the granulation. It has beendiscovered that the incoming particle size of the active ingredient alsoeffects the ultimate particle size distribution of the dry milledagglomerates formed during granulations. For a fixed water quantity, acoarser distribution will result in a finer size distribution of the drymilled agglomerates. Too fine a granulation distribution can lead topoor granulation flow and poor control of individual tablet weightduring the compression step. Thus the narrow particle size constraintspreviously mentioned have also resulted in making the process moreamenable to automation by reducing the variations in water requiredduring the granulation step and producing dry milled granules of theappropriate distribution to prevent the rejection of tablets duringcompression due to unacceptable tablet weight.

The present invention also provides methods of use in inhibitingcompounds of Formula I. Such uses include inhibiting osteoporosis,treating or prevent breast cancer, inhibiting uterine fibrosis,inhibiting endometriosis, and lowering serum cholesterol.

As used herein, the term “effective amount” means an amount of compoundof formula I which is capable of alleviating the symptoms of the variouspathological conditions herein described. The specific dose of acompound administered according to this invention will, of course, bedetermined by the particular circumstances surrounding the caseincluding, for example, the compound administered, the route ofadministration, the state of being of the patient, and the pathologicalcondition being treated. A typical daily dose will contain a nontoxicdosage level of from about 10.0 mg to about 1000 mg/day of a compound ofthe present invention. Preferred daily doses generally will be fromabout 50 mg to about 150 mg/day.

Besides the hydrochloride salt, the compounds of this invention formpharmaceutically acceptable acid and base addition salts with a widevariety of organic and inorganic acids and bases and include thephysiologically acceptable salts which are often used in pharmaceuticalchemistry. Such salts are also part of this invention. Typical inorganicacids used to form such salts include hydrobromic, hydroiodic, nitric,sulfuric, phosphoric, hypophosphoric and the like. Salts derived fromorganic acids, such as aliphatic mono and dicarboxylic acids, phenylsubstituted alkanoic acids, hydroxyalkanoic and hydroxyalkandioic acids,aromatic acids, aliphatic and aromatic sulfonic acids, may also be used.Such pharmaceutically acceptable salts thus include acetate,phenylacetate, trifluoroacetate, acrylate, ascorbate, benzoate,chlorobenzoate, dinitrobenzoate, hydroxybenzoate, methoxybenzoate,methylbenzoate, o-acetoxybenzoate, naphthalene-2-benzoate, bromide,isobutyrate, phenylbutyrate, β-hydroxybutyrate, butyne-1,4-dioate,hexyne-1,4-dioate, caprate, caprylate, chloride, cinnamate, citrate,formate, fumarate, glycollate, heptanoate, hippurate, lactate, malate,maleate, hydroxymaleate, malonate, mandelate, mesylate, nicotinate,isonicotinate, nitrate, oxalate, phthalate, teraphthalate, phosphate,monohydrogenphosphate, dihydrogenphosphate, metaphosphate,pyrophosphate, propiolate, propionate, phenylpropionate, salicylate,sebacate, succinate, suberate, sulfate, bisulfate, pyrosulfate, sulfite,bisulfite, sulfonate, benzene-sulfonate, p-bromophenylsulfonate,chlorobenzenesulfonate, ethanesulfonate, 2-hydroxyethanesulfonate,methanesulfonate, naphthalene-1-sulfonate, naphthalene-2-sulfonate,p-toluenesulfonate, xylenesulfonate, tartarate, and the like. Of course,the preferred salt is the hydrochloride salt.

The pharmaceutically acceptable acid addition salts are typically formedby reacting a compound of formula I with an equimolar or excess amountof acid. The reactants are generally combined in a mutual solvent suchas diethyl ether or benzene. The salt normally precipitates out ofsolution within about one hour to 10 days and can be isolated byfiltration or the solvent can be stripped off by conventional means.

Bases commonly used for formation of salts include ammonium hydroxideand alkali and alkaline earth metal hydroxides, carbonates, as well asaliphatic and primary, secondary and tertiary amines, aliphaticdiamines. Bases especially useful in the preparation of addition saltsinclude ammonium hydroxide, potassium carbonate, methylamine,diethylamine, ethylene diamine and cyclohexylamine.

The compounds of this invention can be administered by a variety ofroutes including oral, rectal, transdermal, subucutaneus, intravenous,intramuscular, and intranasal. These compounds preferably are formulatedprior to administration, the selection of which will be decided by theattending physician. Thus, another aspect of the present invention is apharmaceutical composition comprising an effective amount of a compoundof Formula I, or a pharmaceutically acceptable salt thereof, optionallycontaining an effective amount of estrogen or progestin, and apharmaceutically acceptable carrier, diluent, or excipient.

The total active ingredients in such formulations comprises from 0.1% to99.9% by weight of the formulation. By “pharmaceutically acceptable” itis meant the carrier, diluent, excipients and salt must be compatiblewith the other ingredients of the formulation, and not deleterious tothe recipient thereof.

Pharmaceutical formulations of the present invention can be prepared byprocedures known in the art using well known and readily availableingredients. For example, the compounds of formula I, with or without anestrogen or progestin compound, can be formulated with commonexcipients, diluents, or carriers, and formed into tablets, capsules,suspensions, powders, and the like. Examples of excipients, diluents,and carriers that are suitable for such formulations include thefollowing: fillers and extenders such as starch, sugars, mannitol, andsilicic derivatives; binding agents such as carboxymethyl cellulose andother cellulose derivatives, alginates, gelatin, andpolyvinyl-pyrrolidone; moisturizing agents such as glycerol;disintegrating agents such as calcium carbonate, sodium bicarbonate andcross-linked povidone (cross povidone); agents for retarding dissolutionsuch as paraffin; resorption accelerators such as quaternary ammoniumcompounds; surface active agents such as cetyl alcohol, polysorbate 80,glycerol monostearate; adsorptive carriers such as kaolin and bentonite;and lubricants such as talc, calcium and magnesium stearate, and solidpolyethyl glycols.

The compounds also can be formulated as elixirs or solutions forconvenient oral administration or as solutions appropriate forparenteral administration, for example, by intramuscular, subcutaneousor intravenous routes. Additionally, the compounds are well suited toformulation as sustained release dosage forms and the like. Theformulations can be so constituted that they release the activeingredient only or preferably in a particular physiological location,possibly over a period of time. The coatings, envelopes, and protectivematrices may be made, for example, from polymeric substances or waxes.

Compounds of formula I, alone or in combination with anotherpharmaceutical agent, generally will be administered in a convenientformulation. The following formulation examples only are illustrativeand are not intended to limit the scope of the present invention.

Formulations

In the formulations which follow, “Raloxifene” means a compound offormula I, including salts and solvates thereof.

Formulation 1: Gelatin Capsules

Hard gelatin capsules are prepared using the following:

Ingredient Quantity (mg/capsule) Raloxifene 10.0-1000  Starch, NF 0-650Starch flowable powder 0-650 Silicone fluid 350 centistokes 0-15 

The formulation above may be changed in compliance with the reasonablevariations provided.

A tablet formulation is prepared using the ingredients below:

Formulation 2: Tablets

Ingredient Quantity (mg/tablet) Raloxifene  2.5-1000 Cellulose,microcrystalline 200-650 Silicon dioxide, fumed  10-650 Stearic acid 5-15The components are blended and compressed to form tablets.

Alternatively, tablets each containing 2.5-1000 mg of Raloxifene aremade up as follows:

Formulation 3: Tablets

Ingredient Quantity (mg/tablet) Raloxifene 25-1000 Starch 45 Cellulose,microcrystalline 35 Polyvinylpyrrolidone 4 (as 10% solution in water)Sodium carboxymethyl cellulose 4.5 Magnesium stearate 0.5 Talc 1

Raloxifene, starch, and cellulose are passed through a No. 45 mesh U.S.sieve and mixed thoroughly. The solution of polyvinylpyrrolidone ismixed with the resultant powders which are then passed through a No. 14mesh U.S. sieve. The granules so produced are dried at 50°-60° C. andpassed through a No. 18 mesh U.S. sieve. The sodium carboxymethylstarch, magnesium stearate, and talc, previously passed through a No. 60U.S. sieve, are then added to the granules which, after mixing, arecompressed on a tablet machine to yield tablets.

SuspensiOns each containing 0.1-1000 mg of medicament per 5 ml dose aremade as follows:

Formulation 4: Suspensions

Ingredient Quantity (mg/5 ml) Raloxifene 0.1-1000 mg Sodiumcarboxymethyl cellulose 50 mg Syrup 1.25 mg Benzoic acid solution 0.10mL Flavor q.v. Color q.v. Purified water to 5 mLThe medicament is passed through a No. 45 mesh U.S. sieve and mixed withthe sodium carboxymethyl cellulose and syrup to form a smooth paste. Thebenzoic acid solution, flavor, and color are diluted with some of thewater and added, with stirring. Sufficient water is then added toproduce the required volume.An aerosol solution is prepared containing the following ingredients:

Formulation 5: Aerosol

Ingredient Quantity (% by weight ) Raloxifene 0.25 Ethanol 25.75Propellant 22 70.00 (Chlorodifluoromethane)

Raloxifene is mixed with ethanol and the mixture added to a portion ofthe propellant 22, cooled to 30° C., and transferred to a fillingdevice. The required amount is then fed to a stainless steel containerand diluted with the remaining propellant. The valve units are thenfitted to the container.

Suppositories are prepared as follows:

Formulation 6: Suppositories

Ingredient Quantity (mg/suppository) Raloxifene 250 Saturated fatty acidglycerides 2,000

Raloxifene is passed through a No. 60 mesh U.S. sieve and suspended inthe saturated fatty acid glycerides previously melted using the minimalnecessary heat. The mixture is then poured into a suppository mold ofnominal 2 g capacity and allowed to cool.

An intravenous formulation is prepared as follows:

Formulation 7: Intravenous Solution

Ingredient Quantity Raloxifene 50 mg Isotonic saline 1,000 mL

The solution of Raloxifene is intravenously administered to a patient ata rate of about 1 mL per minute.

Formulation 8: Combination Capsule I

Ingredient Quantity (mg/capsule) Raloxifene 50 Premarin 1 Avicel pH. 10150 Starch 1500 117.50 Silicon Oil 2 Tween 80 0.50 Cab-O-Sil 0.25

Formulation 9: Combination Capsule II

Ingredient Quantity (mg/capsule) Raloxifene 50 Norethylnodrel 5 AvicelpH 101 82.50 Starch 1500 90 Silicon Oil 2 Tween 80 0.50

Formulation 10: Combination Tablet

Ingredient Quantity (mg/capsule) Raloxifene 50 Premarin 1 Corn Starch NF50 Povidone, K29-32 6 Avicel pH 101 41.50 Avicel pH 102 136.50Crospovidone XL10 2.50 Magnesium Stearate 0.50 Cab-O-Sil 0.50

Formulation 11:

Ingredient Quantity (mg/capsule) Raloxifene HCl 60-150Polyvinylpyrrolidone 15.75 Polysorbate 80 5.25 Lactose Anhydrous 264.62Cross-linked 31.5 polyvinylpyrrolidone Stearic Acid 5.25 MagnesiumStearate 2.63

The mixture of raloxifene HCl, lactose, and a portion of thecross-linked polyvinylpyrrolidone is granulated with an aqueous solutionof the polyvinylpyrrolidone and polysorbate 80. The granules are dried,reduced to a suitable size, and mixed with stearic acid, magnesiumstearate, and remaining cross-linked polyvinylpyrrolidone. The mixtureis compressed into individual tablets.

Formulation 12:

Ingredient Quantity (mg/capsule) Raloxifene HCl 60-150Polyvinylpyrrolidone 15.75 Polysorbate 80 5.75 Lactose Anhydrous 132.06Dextrose 132.06 Cross-linked 31.5 polyvinylpyrrolidone Stearic Acid 5.25Magnesium Stearate 2.63

The mixture of raloxifene HCl, lactose anhydrous, dextrose, and aportion of the cross-linked polyvinylpyrrolidone is granulated with analcoholic solution of polyvinylpyrrolidone and polysorbate 80. Thegranules are dried, reduced to a suitable size, and mixed with magnesiumstearate, stearic acid, and remaining cross-linked polyvinylpyrrolidone.The mixture is compressed into individual tablets.

Formulation 13:

Ingredient Quantity (mg/capsule) Raloxifene HCl 60-150 HydroxypropylCellulose 16.00 Sodium Laurylsulfate 10.00 Dextrose 154.00 Cross-linkedsodium 16.00 carboxymethylcellulose Magnesium Stearate 4.00

The mixture of raloxifene HCl, dextrose, and cross-linked sodiumcarboxymethylcellulose is granulated with an aqueous solution ofhydroxypropyl cellulose and sodium laurylsulfate. The granules aredried, reduced to a suitable size, and mixed with magnesium stearate.The mixture is compressed into individual tablets.

Formulation 14:

Ingredient Quantity (mg/capsule) Raloxifene HCl 30.00 Lactose Anhydrous144.00 Lactose, Hydrous spray 36.00 Dried Polyvinylpyrrolidone 12.00Polysorbate 80 2.40 Cross-linked 14.40 polyvinylpyrrolidone MagnesiumStearate 1.20

The mixture of raloxifene HCl, lactose anhydrous, spray-dried hydrouslactose, and a portion of the cross-linked polyvinylpyrrolidone isgranulated with an aqueous solution of polyvinylpyrrolidone andpolysorbate 80. The granules are dried, reduced to a suitable size, andmixed with magnesium stearate and remaining cross-linkedpolyvinylpyrrolidone. The mixture is compressed into individual tabletsyielding a tablet weight of 240 mg.

Formulation 15:

Ingredient Quantity (mg/capsule) Raloxifene HCl 30.00 Lactose Anhydrous160.00 Hydroxypropyl Cellulose 11.00 Poloxamer 7.00 Cross-linked sodium23.00 carboxymethylcellulose Stearic Acid 2.00 Magnesium Stearate 4.00

The mixture of raloxifene HCl, anhydrous lactose, and cross-linkedsodium carboxymethylcellulose is granulated with an aqueous solution ofpoloxamer and hydroxypropyl cellulose. The granules are dried, reducedto a suitable size, and mixed with stearic acid and magnesium stearate.The mixture is then compressed into individual tablets yielding a tabletweight of 240 mg.

Formulation 16:

Ingredient Quantity (mg/capsule) Raloxifene HCl 30.00 Lactose 89.00Dextrose 89.00 Hydroxypropyl 10.00 methylcellulose Sodium Laurylsulfate5.00 Cross-linked sodium 12.00 polyvinylpyrrolidone Stearic Acid 5.00

The mixture of raloxifene HCl, lactose, dextrose, and cross-linkedpolyvinylpyrrolidone is granulated with an aqueous solution ofhydroxypropyl methylcellulose and sodium laurylsulfate. The granules aredried, reduced to a suitable size, and mixed with the stearic acid. Themixture is then compressed into individual tablets yielding a tabletweight of 240 mg.

Formulation 17:

Ingredient Quantity (mg/capsule) Raloxifene HCl 60.00 Lactose Anhydrous156.00 Polyvinylpyrrolidone 7.20 Polysorbate 80 7.20 Cross-linked sodium7.20 polyvinylpyrrolidone Magnesium Stearate 2.40

The mixture of raloxifene HCl, lactose anhydrous, and cross-linkedpolyvinylpyrrolidone is granulated with an aqueous solution ofpolyvinylpyrrolidone and polysorbate 80. The granules are dried, reducedto a suitable size, and mixed with magnesium stearate. The mixture isthen compressed into individual tablets yielding a tablet weight of 240mg.

Formulation 18:

Ingredient Quantity (mg/capsule) Raloxifene HCl 60.00 Lactose Anhydrous120.00 Lactose, hydrous spray- 30.00 dried Polyvinylpyrrolidone 12.00Polysorbate 80 2.40 Cross-linked sodium 14.40 polyvinylpyrrolidoneMagnesium Stearate 1.20

The mixture of raloxifene HCl, lactose anhydrous, spray-dried hydrouslactose, and a portion of the cross-linked polyvinylpyrrolidone isgranulated with an aqueous solution of polyvinylpyrrolidone andpolysorbate 80. The granules are dried, reduced to a suitable size, andmixed with magnesium stearate and remaining cross-linkedpolyvinylpyrrolidone. The mixture is then compressed into individualtablets yielding a tablet weight of 240 mg.

Formulation 19:

Ingredient Quantity (mg/capsule) Raloxifene HCl 60.00 Mannitol 77.00Dextrose 73.00 Hydroxypropyl 7.00 methylcellulose Polysorbate 80 4.00Sodium Starch Glycolate 14.00 Stearic Acid 4.00 Magnesium Stearate 1.00

The mixture of raloxifene HCl, mannitol, dextrose, and sodium starchglycolate is granulated with an aqueous solution of polysorbate 80 andhydroxypropyl methylcellulose. The granules are dried, reduced to asuitable size, and mixed with stearic acid and magnesium stearate. Themixture is then compressed into individual tablets yielding a tabletweight of 240 mg.

Formulation 20:

Ingredient Quantity (mg/capsule) Raloxifene HCl 150.00 Lactose Anhydrous41.00 Lactose, hydrous spray- 10.25 dried Polyvinylpyrrolidone 11.50Polysorbate 80 2.30 Cross-linked sodium 13.80 polyvinylpyrrolidoneMagnesium Stearate 1.15

The mixture of raloxifene HCl, anhydrous lactose, hydrous spray-driedlactose, and a portion of the cross-linked polyvinylpyrrolidone isgranulated with an aqueous solution of polyvinylpyrrolidone andpolysorbate 80. The granules are dried, reduced to a suitable size, andmixed with magnesium stearate and the remaining cross-linkedpolyvinylpyrrolidone. The mixture is then compressed into individualtablets yielding a tablet weight of 230 mg.

Formulation 21:

Ingredient Quantity (mg/capsule) Raloxifene HCl 150.00 Lactose, hydrousspray- 56.00 dried Polyvinylpyrrolidone 7.00 Polysorbate 80 1.20Cross-linked sodium 13.80 polyvinylpyrrolidone Magnesium Stearate 2.00

The mixture of raloxifene HCl, hydrous spray-dried lactose, and aportion of the cross-linked polyvinylpyrrolidone is granulated with anaqueous solution of polyvinylpyrrolidone and polysorbate 80. Thegranules are dried, reduced to a suitable size and mixed with magnesiumstearate and remaining cross-linked polyvinylpyrrolidone. The mixture isthen compressed into individual tablets yielding a tablet weight of 230mg.

Formulation 22:

Ingredient Quantity (mg/capsule) Raloxifene HCl 150.00 Lactose,anhydrous 52.40 Polyvinylpyrrolidone 11.50 Polysorbate 80 4.60Polyethylene Glycol 8000 11.50

The mixture of raloxifene HCl and anhydrous lactose is granulated withan aqueous solution of polysorbate 80 and polyvinylpyrrolidone. Thegranules are dried, reduced to a suitable size, and mixed with thepolyethylene glycol 8000. The mixture is then compressed into individualtablets yielding a tablet weight of 230 mg.

Capsules may be prepared using the ingredients and procedures asdescribed below:

Formulation 2:

Ingredient Quantity (mg/capsule) Raloxifene HCl 30.00 Lactose, hydrousspray-dried 178.30 Sodium laurylsulfate 4.60 Cross-linked 9.20polyvinylpyrrolidone Hydroxypropyl 6.90 methylcellulose ColloidalSilicon Dioxide 1.00

The mixture of raloxifene HCl, hydrous spray-dried lactose, andcross-linked polyvinylpyrrolidone is granulated with an aqueous solutionof sodium laurylsulfate and hydroxypropyl methylcellulose. The granulesare dried, reduced to a suitable size, and mixed with colloidal silicondioxide. This mixture is then filled into Size 3 hard-shell gelatincapsules utilizing conventional encapsulating equipment, with eachcapsule containing 230 mg of the final mixture.

Formulation 24:

Ingredient Quantity (mg/capsule) Raloxifene HCl 60.00 Lactose, hydrousspray-dried 148.30 Sodium laurylsulfate 4.60 Cross-linked 9.20polyvinylpyrrolidone Hydroxypropyl 6.90 methylcellulose ColloidalSilicon Dioxide 1.00

The mixture of raloxifene HCl, hydrous spray-dried lactose, andcross-linked polyvinylpyrrolidone is granulated with an aqueous solutionof sodium laurylsulfate and hydroxypropyl methylcellulose. The granulesare dried, reduced to a suitable size, and mixed with colloidal silicondioxide. This mixture is then filled into Size 3 hard-shell gelatincapsules utilizing conventional encapsulating equipment, with eachcapsule containing 230 mg of the final mixture.

Formulation 25:

Ingredient Quantity (mg/capsule) Raloxifene HCl 150.00 Lactose, hydrousspray-dried 58.30 Sodium laurylsulfate 4.60 Cross-linked 9.20polyvinylpyrrolidone Hydroxypropyl 6.90 methylcellulose ColloidalSilicon Dioxide 1.00

The mixture of raloxifene HCl, hydrous spray-dried lactose, andcross-linked polyvinylpyrrolidone is granulated with an aqueous solutionof sodium laurylsulfate and hydroxypropyl methylcellulose. The granulesare dried, reduced to a suitable size, and mixed with colloidal silicondioxide. This mixture is then filled into Size 3 hard-shell gelatincapsules utilizing conventional encapsulating equipment, with eachcapsule containing 230 mg of the final mixture.

1. A compound of formula I

and pharmaceutically acceptable salts and solvates thereof,characterized in that the compound is in particulate form, saidparticles having a mean particle size of less than about 25 microns. 2.The compound of claim 1 wherein said particles have a mean particle sizeof between about 5 and about 20 microns.
 3. The compound of claim 2wherein at least about 90% of said particles have a size of less thanabout 50 microns.
 4. The compound of claim 3 wherein at least 90% ofsaid particles have a size of less than about 35 microns.
 5. A compoundof claim 1 which is the non-solvated crystalline6-hydroxy-2-(4-hydroxy-phenyl)-3-[4-(2-piperidinoethoxy)benzoyl]benzo[b]thiophenehydrochloride having substantially the following X-ray diffractionpattern obtained with copper radiation: d-line spacing I/I_(o)(Angstroms) (×100) 13.3864 71.31 9.3598 33.16 8.4625 2.08 7.3888 7.576.9907 5.80 6.6346 51.04 6.1717 29.57 5.9975 5.67 5.9135 9.87 5.646738.47 5.4773 10.54 5.2994 4.74 4.8680 4.03 4.7910 5.98 4.6614 57.504.5052 5.75 4.3701 9.03 4.2516 69.99 4.2059 57.64 4.1740 65.07 4.081912.44 3.9673 22.53 3.9318 100.00 3.8775 9.07 3.7096 33.38 3.6561 21.653.5576 3.36 3.5037 7.97 3.4522 18.02 3.4138 4.65 3.2738 10.23 3.18578.90 3.1333 6.24 3.0831 9.43 3.0025 12.13 2.9437 4.96 2.8642 7.70 2.790411.95 2.7246 3.05 2.6652 3.32 2.5882 7.30


6. A pharmaceutical formulation comprising or formulated using thecompound of claim 5 and one or more pharmaceutically acceptablecarriers, diluents, or excipients.
 7. A pharmaceutical compositioncomprising or formulated using a compound according to claim 1, or apharmaceutically acceptable salt or solvate thereof, in combination withone or more pharmaceutically acceptable carriers, diluents orexcipients.
 8. A method of inhibiting osteoporosis comprisingadministering an effective amount of a compound of formula I to a personin need thereof.
 9. A method of lowering serum lipid levels comprisingadministering a compound of claim 1, or a pharmaceutically acceptablesalt or solvate thereof, to a person in need thereof.
 10. A method forpreventing breast cancer comprising administering to a woman in needthereof an effective amount of a compound of claim 1, or apharmaceutically acceptable salt or solvate thereof.
 11. An article ofmanufacture comprising packaging material containing a pharmaceuticalformulation, said packaging material further containing labellingindicating said pharmaceutical formulation is useful for inhibiting ahuman pathology, said pharmaceutical formulation comprising orformulated using a compound of claim
 1. 12. A compound of formula I

and pharmaceutically acceptable salts and solvates thereof,characterized in that the compound is in particulate form, at leastabout 90% of said particles having a particle size of less than about 50microns.
 13. The compound of claim 12 wherein at least about 90% of saidparticles have a size of less than 35 microns.
 14. The compound of claim13 wherein said particles have a mean particle size of less than about25 microns.
 15. The compound of claim 14 wherein said particles have amean particles size of between about 5 and about 20 microns.
 16. Acompound of claim 12 which is the non-solvated crystalline6-hydroxy-2-(4-hydroxy-phenyl)-3-[4-(2-piperidinoethoxy)benzoyl]benzo[b]thiophenehydrochloride, having substantially the following X-ray diffractionpattern obtained with copper radiation: d-line spacing I/I_(o)(Angstroms) (×100) 13.3864 71.31 9.3598 33.16 8.4625 2.08 7.3888 7.576.9907 5.80 6.6346 51.04 6.1717 29.57 5.9975 5.67 5.9135 9.87 5.646738.47 5.4773 10.54 5.2994 4.74 4.8680 4.03 4.7910 5.98 4.6614 57.504.5052 5.75 4.3701 9.03 4.2516 69.99 4.2059 57.64 4.1740 65.07 4.081912.44 3.9673 22.53 3.9318 100.00 3.8775 9.07 3.7096 33.38 3.6561 21.653.5576 3.36 3.5037 7.97 3.4522 18.02 3.4138 4.65 3.2738 10.23 3.18578.90 3.1333 6.24 3.0831 9.43 3.0025 12.13 2.9437 4.96 2.8642 7.70 2.790411.95 2.7246 3.05 2.6652 3.32 2.5882 7.30


17. A pharmaceutical formulation comprising or formulated using thecompound of claim 16, and one or more pharmaceutically acceptablecarriers, diluents, or excipients.
 18. A pharmaceutical compositioncomprising or formulating using a compound according to claim 12, or apharmaceutically acceptable salt or solvate thereof, with one or morepharmaceutically acceptable carriers, diluents or excipients.
 19. Amethod of inhibiting osteoporosis comprising administering to a personin need thereof an effective amount of a compound of claim 12, or apharmaceutically acceptable salt or solvate thereof.
 20. A method oflowering serum lipid levels, comprising administering to a person inneed thereof an effective amount of a compound of claim 12, or apharmaceutically acceptable salt or solvate thereof.
 21. A method forpreventing breast cancer comprising administering to a woman in needthereof an effective amount of a compound of claim 12, or apharmaceutically acceptable salt or solvate thereof.
 22. An article ofmanufacture comprising packaging material containing a pharmaceuticalformulation, said packaging material further containing labellingindicating said pharmaceutical formulation is useful for inhibiting ahuman pathology, said pharmaceutical formulation comprising orformulated using a compound of claim
 12. 23. A compound of formula I

and pharmaceutically acceptable salts and solvates thereof,characterized in that the compound is in particulate form, saidparticles having a mean particle size of between about 5 and about 20microns, at least about 90% of said particles having a size of less thanabout 35 microns.
 24. A compound according to claim 23 wherein it is thehydrochloride salt thereof.
 25. A compound of claim 23 which is thenon-solvated crystalline6-hydroxy-2-(4-hydroxy-phenyl)-3-[4-(2-piperidinoethoxy)benzoyl]benzo[b]thiophenehydrochloride, having substantially the following x-ray diffractionpattern obtained with copper radiation: d-line spacing I/I_(o)(Angstroms) (×100) 13.3864 71.31 9.3598 33.16 8.4625 2.08 7.3888 7.576.9907 5.80 6.6346 51.04 6.1717 29.57 5.9975 5.67 5.9135 9.87 5.646738.47 5.4773 10.54 5.2994 4.74 4.8680 4.03 4.7910 5.98 4.6614 57.504.5052 5.75 4.3701 9.03 4.2516 69.99 4.2059 57.64 4.1740 65.07 4.081912.44 3.9673 22.53 3.9318 100.00 3.8775 9.07 3.7096 33.38 3.6561 21.653.5576 3.36 3.5037 7.97 3.4522 18.02 3.4138 4.65 3.2738 10.23 3.18578.90 3.1333 6.24 3.0831 9.43 3.0025 12.13 2.9437 4.96 2.8642 7.70 2.790411.95 2.7246 3.05 2.6652 3.32 2.5882 7.30


26. A pharmaceutical formulation comprising or formulated using thecompound of claim 25 and one or more pharmaceutically-acceptablecarriers, diluents, or excipients.
 27. A pharmaceutical compositioncomprising or formulated using a compound according to claim 23, or apharmaceutically acceptable salt or solvate thereof, with one or morepharmaceutically acceptable carriers, diluents or excipients.
 28. Amethod for inhibiting osteoporosis comprising administering to a personin need thereof an effective amount of a compound of claim 23, or apharmaceutically acceptable salt or solvate thereof.
 29. A method oflowering serum lipid levels, comprising administering an effectiveamount of a compound of claim 23, or a pharmaceutically acceptable saltor solvate thereof, to a person in need thereof.
 30. A method forpreventing breast cancer comprising administering to a woman in needthereof an effective amount of a compound of claim 23, or apharmaceutically acceptable salt or solvate thereof.
 31. An article ofmanufacture comprising packaging material containing a pharmaceuticalformulation, said packaging material further containing labellingindicating said pharmaceutical formulation is useful for inhibiting ahuman pathology, said pharmaceutical formulation comprising orformulated using a compound of claim 23.